Objective To establish a rapid, sensitive and specific method for detection of Listeria monocytogenes(Lm) in the import and export foods.Method A pair of oligonucleotide primers and a probe were designed with listeriolysion A (hlyA) gene as target sequence DNA, cell, plasmid of Lm were amplified by real-time PCR technique.Results This trial established DNA standard curve, cell standard curve and plasmid standard curve by real-time PCR technique. The results obtained by the three standard curves were mainly the same.Conclusion Real-time PCR can be used as a sensitive, specific and accurate method for detection of Lm. The inspection system developed in this experiment can be used to do Listeria monocytogenes inspection work in the department of inspection and quarantine.